KR20100122773A - Process for separating and purifying succinic acid from fermentation broth - Google Patents

Abstract

PURPOSE: A method for purifying and isolating succinic acid from fermentation liquid using strain is provided to generate crystalline succinic acid and salt having high solubility. CONSTITUTION: A method for isolating and purifying succinic acid from fermentation liquid using a strain comprises: a step of adding base to fermentation liquid to generate precipitate containing succinic acid; and a step of adding hydrochloric acid solution, nictric acid solution or mixture solution to the precipitate to generation crystalline succinic acid. The method for purifying comprises a step of heating the fermentation liquid or supplying air or nitrogen to remove CO_3^2- contained in the fermentation liquid.

Description

PROCESS FOR SEPARATING AND PURIFYING SUCCINIC ACID FROM FERMENTATION BROTH}

The present invention relates to a method for separating and purifying succinic acid from a fermentation broth prepared with succinic acid using a strain.

Recently, interest in developing eco-friendly chemical products using biological resources that can reduce the dependence on limited fossil resources and use renewable and sustainable materials has been rapidly increasing. Among the compounds that can be derived from biological sources replacing fossil raw materials, succinic acid is the platform of C4 compounds, such as BDO (1,4-butanediol), THF (tetrahydrofuran), GBL (gamma-butyrolactone), It is expected to be used as a raw material for the production of general-purpose products such as n-methylpyrrolidone (NMP). In addition, the process of producing succinic acid from glucose is significant because it can be an active countermeasure to the Kyoto Convention on the reduction of greenhouse gas emissions through fixed carbon dioxide fermentation.

Succinic acid has been widely used as a raw material in the food, pharmaceutical and cosmetic industries. Generally, succinic acid is made from normal butane (n-butane) via maleic anhydride. However, succinic acid can be prepared through fermentation as well as this petrochemical process, as a metabolite of the TriCarboxylic Acid Cycle (TCA) cycle, which is one of the main metabolic pathways of organisms.

However, in the fermentation broth formed by fermenting carbohydrates using the strain, in addition to succinic acid, which is a desired product, other organic acids such as dermic acid, citric acid, oxal acetic acid, formic acid, acetic acid, and fumaric acid, K ⁺ , NH ₄ ⁺ , Mg ^{2 +} , Na ⁺ cation, such _{^{as, CO 3 2 -, PO 4}} -, SO 4 2 - is the anion with the unreacted carbohydrate, amino acid, such as containing a large amount of water. Therefore, in order to obtain high purity succinic acid from the fermentation broth, it is necessary to efficiently remove the above-mentioned substances from the fermentation broth.

However, the separation and purification of high purity succinic acid according to the processes known to the art up to now requires a lot of energy, and due to the high cost of the separation and purification of succinic acid, which accounts for about 50 to 70% of the total production cost, The production cost of succinic acid is significantly higher than that of existing petrochemical processes.

Separation and purification of succinic acid from fermentation broth known to the art to date include electrodialysis-crystallization, ion exchange resin-crystallization, precipitation-crystallization and extraction-crystallization.

The electrodialysis-crystallization method or the ion exchange resin-crystallization method includes the steps of replacing succinate in the fermentation broth with succinic acid using an ion exchange membrane or an ion exchange resin; And selectively separating succinic acid having relatively low solubility by evaporating water in the aqueous succinic acid solution. This method has the advantages of rapid treatment and non-productive production. However, the ion exchange membrane and the ion exchange resin used are expensive, and the long term use of the ion exchange membrane and the ion exchange resin is contaminated, resulting in poor effectiveness, and when applied to a commercial production scale, there is a problem of low operational efficiency. . In addition, electrodialysis-crystallization or ion-exchange resin-crystallization has not been commercialized due to the high energy and high cost of water evaporation.

The extraction-crystallization method includes a step of selectively removing other organic acids from fermentation broth using an extraction solvent; And selectively separating succinic acid by evaporating water from the fermentation broth from which other organic acids have been removed. This method also has high energy and high cost problems due to the water evaporation process, and also has problems such as high cost of the extraction solvent, recycling of the organic solvent, low selectivity of the organic solvent.

The precipitation-crystallization method includes the steps of selectively precipitated succinic acid in the fermentation broth using calcium hydroxide; And an aqueous solution of sulfuric acid is added to the precipitate to separate insoluble calcium sulfate from succinic acid in aqueous solution and to crystallize succinic acid by evaporating water from the separated succinic acid in aqueous solution. This method also has a problem of high energy and cost according to the water evaporation process, and also has a problem of generating a large amount of calcium sulfate as a solid waste.

The present inventors added calcium chloride instead of a low solubility salt such as calcium sulfate as a by-product when adding an aqueous hydrochloric acid solution and / or an aqueous nitric acid solution to a succinate obtained by adding a base to a fermentation broth prepared with succinic acid using a strain. Since salts with high solubility, such as calcium nitrate and the like, are produced, it has been found that high purity succinic acid in crystalline state can be obtained without an additional purification step. The present invention is based on this.

The present invention is a method for separating and purifying succinic acid from a fermentation broth prepared with succinic acid using a strain, wherein a base is added to a fermentation broth with or expressing succinic acid using a strain to produce a precipitate containing succinate. step; And adding an acidic aqueous solution selected from an aqueous hydrochloric acid solution, an aqueous nitric acid solution, and a mixture thereof to the precipitate containing the succinic acid salt to produce succinic acid in a crystalline state.

The present invention is a method for separating and purifying succinic acid from a fermentation broth, wherein an aqueous hydrochloric acid solution and / or an aqueous nitric acid solution is added to a precipitate containing succinate produced by adding a base to a fermentation broth in which succinic acid is expressed or expressed using a strain. As a result, salts with high solubility are produced together with succinic acid in the crystalline state, so that high purity succinic acid can be obtained even by using a simple low-cost filtration device.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

In the present invention, the first supernatant means a liquid substance other than a precipitate containing succinate obtained by adding a base to a fermentation broth in which succinic acid is expressed or expressed.

In the present invention, the second supernatant means a liquid substance other than succinic acid in the crystal state obtained by adding an aqueous hydrochloric acid solution or an aqueous nitric acid solution to a precipitate containing succinate.

The present invention is characterized by obtaining a succinic acid in a low cost and high efficiency by adding an aqueous solution of hydrochloric acid and / or an aqueous solution of nitric acid to a precipitate containing a succinate produced by adding a base to the fermentation broth expressed succinic acid using a strain. It is done.

In general, when a strain such as glucose or carbohydrate is decomposed by the strain, in addition to succinic acid, other organic acids such as dermic acid, citric acid, oxal acetic acid, formic acid, acetic acid and fumaric acid, K ⁺ , NH ₄ ⁺ , Mg ^{2 +} , Na ⁺ cations, such _{^{as, CO 3 2 -, PO 4}} -, SO 4 2 - yi fermentation liquor containing anion, the unreacted glucose, or carbohydrates, amino acids with an excess of water, such as is formed. In order to obtain the desired high purity succinic acid from the fermentation broth, it should be possible to selectively separate only succinic acid from the fermentation broth.

Thus, in the conventional precipitation-crystallization method, in order to separate and purify succinic acid from the fermentation broth, a base (ex. Calcium hydroxide) is added to the fermentation broth to generate succinate precipitate (ex. Calcium succinate), and then to the succinate precipitate. Dilute sulfuric acid solution was added. At this point, succinic acid (solubility 74.5 g / l at room temperature), calcium sulfate (2.4 g / l at room temperature) and other by-products (other organic acids, anionic derivatives) are produced, of which calcium sulfate with the lowest solubility is selective. Precipitated to obtain a relatively purified aqueous succinic acid solution. In this case, however, solid waste calcium sulfate (CaSO ₄ ) is precipitated, while other by-products (other organic acids, anion-derived acids) and large amounts of water are included in the supernatant together with succinic acid to obtain succinic acid in crystalline state. In order to solve the problem, an additional separation process was required.

Thus, the present inventors have experimented to add succinic acid (solubility at room temperature: 74.5 g / l), calcium chloride, and / or when adding a high concentration of hydrochloric acid solution and / or high concentration of nitric acid solution to the succinate precipitate instead of dilute sulfuric acid solution Or calcium nitrate (solubility at room temperature: 745 g / l and 1212 g / l, respectively), and other by-products (other organic acids, anionic derivatives) are produced to selectively precipitate the least succinic acid In addition, it was found that only succinic acid in the crystal state could be easily obtained even by using a simple low-cost filtration device.

For example, when a base such as calcium hydroxide is added to a fermentation broth containing a large amount of succinic acid, a succinate (ex.calcium succinate) consisting of a succinate anion and a base cation (ex.Ca ²⁺ ) precipitates. do. Subsequently, when an aqueous hydrochloric acid solution (and / or nitric acid solution) is added to the separated succinate precipitate, the cation that formed the succinate as a reaction by-product together with succinic acid and hydrochloric anion (and / or nitric acid anion in the aqueous hydrochloric acid solution). ) Salts [ex. CaCl ₂ (and / or Ca (NO ₃ ) ₂ )] is produced. In this case, since the resulting calcium chloride and / or calcium nitrate are contained in the supernatant with a higher solubility than succinic acid, succinic acid may be selectively precipitated. In addition, other organic acid derivatives (blood salts, citrates, oxal acetates, formates, acetates, fumarates) and anionic derivatives (calcium sulfate, calcium phosphate, calcium carbonate) contained in succinate precipitates Other organic acids (pibric acid, citric acid, oxalacetic acid, formic acid, acetic acid, fumaric acid) and anionic derivatives (sulfuric acid, phosphoric acid, carbonic acid) produced by reaction with aqueous hydrochloric acid (and / or aqueous nitric acid) are also more soluble than succinic acid. Like calcium chloride (and / or calcium nitrate) it is contained in the supernatant. Therefore, the present invention could easily separate high-purity succinic acid crystals from the supernatant by using a simple low-cost filtration device such as a filter.

As described above, since the present invention can easily obtain crystalline succinic acid using a low-cost filtration device such as a filter, high energy and costly water evaporation to obtain succinic acid in crystalline state, unlike conventional succinic acid separation and purification methods. There is no need to perform the process, which can simplify the overall process for obtaining succinic acid and reduce costs.

In addition, when succinic acid crystals are formed from succinate using a conventional aqueous sulfuric acid solution, insoluble calcium sulfate is produced as a by-product, wherein the produced calcium sulfate is difficult to recycle and can be used only for extremely limited purposes. On the other hand, in the case of the reaction by-products produced in the present invention, such as calcium chloride, it is separated into hydrochloric acid and calcium hydroxide through a simple high temperature hydration reaction, and can be used again in the required stage. Can be.

The present invention is a method for separating and purifying succinic acid from a fermentation broth prepared with succinic acid using a strain, the method comprising a succinate produced by adding a base to the fermentation broth with or expressed succinic acid using a strain To the precipitate, adding an aqueous hydrochloric acid solution and / or an aqueous nitric acid solution to produce succinic acid in crystalline state.

Ⅰ. Sediment formation containing succinate

The precipitate containing succinate used in the present invention can be prepared by the following method, but is not limited thereto.

Ⅰ-1. First method of forming a precipitate containing succinate

The precipitate 60 containing the succinate salt is formed using the strain 20 to form a fermentation broth 30 expressing succinic acid; And adding a base 40 to the fermentation broth 30 in which the succinic acid is expressed, thereby producing a precipitate 60 containing succinate.

(1) Preparation of fermentation broth

The strain 20 for producing succinic acid by fermentation used in the present invention may be used without particular limitation as long as it is a strain capable of expressing succinic acid generally known in the art. Examples of such strains include Anaerobiospirillum succinicproducens, Mannheimia succinicproducens, Escherichia coli, Corynebacterium glutamicumium (Corynebacterium) glutamicum), Actinobacillus succinogens, Brevibacterium flavum, and Zymomonas mobilis, but are not limited thereto.

By metabolizing carbohydrates with nutrients in the presence of carbon dioxide using these strains, a fermentation broth containing a large amount of succinic acid can be obtained. The fermentation broth can contain not only dissociated succinic acid, but also other dissociated organic acids (ex. acetic acid, formic acid, acetic acid, fumaric acid, etc.), a cation (K ^+, NH4 ^+, Mg ^{2 +,} Na ^+), anions _{^{(CO 3 2 -, PO 4}} -, SO 4 2 -) can be obtained by the fermentation broth contains the . In addition to the above substances, the fermentation broth contains not only unreacted carbohydrates, amino acids and excess water, but also used strains.

The formation process of the fermentation broth may be performed in a batch form, a fed-batch form, or a continuous form.

In the present invention, before supplying the fermentation broth 30 as described above to the succinate formation region (I-1 (b)), the strain may be separated from the fermentation broth with the succinic acid expressed using a centrifuge in the centrifugation zone. (Not shown). At this time, by recovering the separated strain and using it again in the fermentation broth forming region, it is possible to reduce the overall cost.

In the present invention, before transfer to the fermentation broth 30 strain is not a / separation separated by acid salt formation region (Ⅰ-1 (b)), optionally impurity anions ^{_{(CO 3 2 -, PO 4}} -, SO ₄ ^2- ), in particular carbonate (CO ₃ ^2- ), may be removed in advance from the fermentation broth in which the succinic acid is expressed. If the fermentation broth contains anions, the fermentation broth is produced when the base (ex. Ca (OH) _2, etc.) added to the fermentation broth later reacts with the dissociated succinic acid in the fermentation broth. The anion also reacts with the base to produce other impurities (ex. CaCO ₃ , Ca (PO ₄ ) ₂ , CaSO _4, etc.), wherein the other impurities are precipitated together with the succinate, which is then used to determine the succinic acid in the crystalline state. This is because the purity can be reduced.

As a method of removing the anion, a method of removing anion contained in the fermentation broth by heating a fermentation broth in which succinic acid is expressed, or supplying air or nitrogen to the fermentation broth in which the succinic acid is expressed using aeration (aeration) By removing the anion contained in the fermentation broth, and the like, but is not limited thereto. In this way, the anions contained in the fermentation broth can be removed in a gaseous state. In particular, carbon dioxide (CO ₃ ^2- ) containing a large amount of carbon dioxide (CO ₃ ^2- ) in the preparation of the fermentation broth expressing succinic acid is carbon dioxide in the gaseous state. Can be removed.

The heating temperature of the fermentation broth is preferably in the range of about 50 to 100 ° C. If the heating temperature of the fermentation broth is less than 50 ° C., the rate of carbonate removal is low. If the heating temperature of the fermentation broth is more than 100 ° C., the energy consumption is high.

However, when the content of the anion in the fermentation broth is small, the anion may be effectively removed through subsequent steps even if the anion removal step is not performed, and thus, it is not necessary to perform the anion removal step.

(2) succinate precipitation

When the base 40 is added while supplying the fermentation broth containing succinic acid 30 to the succinate formation region (I-1 (b)) using the fermentation broth prepared as described above, that is, the strain, the substances in the base and the fermentation broth Reaction produces a precipitate 60 and a supernatant (hereinafter referred to as a 'first supernatant') 50. Here, the precipitate 60 contains a small amount of salts induced by succinate and anion in the form of a precipitate, and the first supernatant 50 contains dissolved unreacted carbohydrates, amino acids, cations, and the like. .

Base 40 that can be used in the present invention is not particularly limited as long as it is a substance capable of selectively precipitate succinic acid in the fermentation broth. Examples of such materials include, but are not limited to, calcium hydroxide (Ca (OH) ₂ ), magnesium hydroxide (Mg (OH) ₂ ), calcium carbonate (CaCO ₃ ), magnesium carbonate (MgCO ₃ ), and mixtures thereof. Do not. Here, among the succinates formed by reacting the base with dissociated succinic acid (succinic anion), calcium succinate (solubility: about 15 g / l) is more precipitated than magnesium succinate (solubility: about 45 g / l). Since it happens well, it is preferable to use calcium hydroxide or calcium carbonate.

The base 40 may be added to the fermentation broth at room temperature. In addition, the present inventors know that when the base is added to the fermentation broth at a higher temperature than at room temperature, the present inventors can obtain a precipitate containing a larger amount of succinate, and it is about 100 ° C. or less, preferably about 50 to 100 ° C. The base 40 was added to the fermentation broth 30 heated or being heated to a high temperature in the range. If the addition of the base to the fermentation broth is performed under a temperature of more than 100 ℃ may be inefficient because a large amount of energy is consumed during the evaporation of water. For example, at room temperature, the concentration of calcium succinate in the supernatant is about 15 g / l, whereas at 80 ° C the concentration of calcium succinate is lowered to about 11 g / l, resulting in higher amounts of calcium succinate in the form of sediments. there was.

The succinate (ex. Calcium succinate, magnesium succinate, etc.) 60 produced by adding the base 40 to the fermentation broth 30 is an unreacted carbohydrate, amino acid, cation or other organic acid (ex. Skin) in the fermentation broth. Citric acid, citric acid, oxal acetic acid, formic acid, acetic acid, fumaric acid, and the like) may be selectively precipitated because of its relatively low solubility in water compared to the induced salts formed by reaction with a base. At this time, the salts induced by the unreacted carbohydrates, amino acids, cations, and other organic acids in the fermentation broth are contained in the first supernatant 50 because of high solubility in water.

However, when the acid salt, such as the precipitation, the inner anion the fermentation broth ^{_{(CO 3 2 -, PO 4}} -, SO 4 2 -) is precipitated by reaction of added base, and unlike any other material, thereby generating Precipitates contain salts induced by anions in addition to succinate. When the precipitate contains salts induced by anions other than succinate, the purity of succinic acid in the final crystal state may be lowered later.

Therefore, in the present invention, the pH of the fermentation broth expressing succinic acid is maintained at about 4.5 to 6.5 or less upon addition of the base so that more precipitate of succinate is formed and less precipitate of salt induced by anion is formed. This is because the reaction of the succinic acid anion-base has a higher selectivity than the reaction of the other anion-base at a low pH. As a result, the formation of salts induced by anions can be minimized to improve the purity of the resulting succinate-containing precipitate, which in turn can improve the quality of the final crystalline succinic acid.

The precipitate 60 and the first supernatant 50 thus formed may be separated from one another via a filtration device in a filtration zone using a filtration device such as a filter, if desired. Thereafter, the precipitate 60 containing the separated succinate is transferred to the succinic acid crystal forming region (II), and at the same time, the first supernatant 50 is recovered and reused in the fermentation broth forming region I-1 (a). Can be.

However, when the concentration of the impurities in the first supernatant is inhibited by strain fermentation when the separated first supernatant 50 is recovered and reused, the impurities in the first supernatant may be purged through a purge process. It is appropriate to reuse the concentration after reducing it below a certain level.

Specifically, when the concentration of impurities in the separated first supernatant is at a level inhibited by strain fermentation in the range of about 1 wt% or more, preferably 1 wt% to 2 wt%, the first supernatant 50 Rather than being transferred directly to the fermentation broth forming region (I-1 (a)) and reused, the first supernatant is further added to the purge region (IV) to add a portion of the first supernatant and discarded. After reducing the concentration of impurities within 1% by weight or less, it is appropriate to reuse in the formation of the fermentation broth.

I-2. Second method of forming a precipitate containing succinate

On the other hand, in order to commercialize the production of succinic acid from biological resources as described above, it is important to produce succinic acid with improved yield in the process of forming the fermentation broth and to obtain succinic acid with improved quality at low cost in the separation and purification process.

However, in the case of the first method of forming a precipitate containing succinate, when the concentration of succinic acid in the fermentation broth increases as the fermentation progresses, the product inhibition phenomenon is inhibited by the growth of the strain by the product succinic acid. It is not possible to obtain more than a certain concentration of succinic acid. In order to solve this problem, a strain having a strong resistance to succinic acid as a product should be used, or the succinic acid produced must be continuously converted or discharged so as not to lower the growth of the strain, thereby keeping the concentration of succinic acid in the fermentation broth low.

Thus, in the present invention, by adding a base to the fermentation broth in which succinic acid is expressed during the fermentation process by the strain to form a precipitate containing succinate, the concentration of succinic acid in the fermentation broth can be kept low, the yield of the final succinic acid This can be improved. Forming a succinate-containing precipitate in this manner is referred to below as a 'second method of forming a succinate-containing precipitate.'

Specifically, in the first method of forming a precipitate containing the succinate described above, after the succinic acid is expressed using a strain to form a fermentation broth, that is, after the fermentation is completed, the base is added to the fermentation broth to form succinate. In the second method of forming a precipitate containing succinate, while forming a precipitate containing succinate, a base is added to the fermentation broth in which succinic acid is expressed during the fermentation process. It is added to form a precipitate containing succinate. In other words, in the first method, the formation of the fermentation broth in which succinic acid is expressed and the formation of the precipitate containing succinate are carried out in separate processes, while the second method is the formation of the fermentation broth in which succinic acid is expressed and the precipitate containing succinate. Is simultaneously formed in one process.

That is, the precipitate 60 containing succinate, as shown in Figure 3, using the strain 20 to form a fermentation broth expressing succinic acid, the base 40 in the fermentation broth in which the succinic acid is expressed It can be formed by the step of forming a precipitate 60 containing succinate.

The strain 20 usable in the present invention can be used without particular limitation as long as it is a strain capable of expressing succinic acid generally known in the art, similarly to the first method of forming the precipitate containing the succinate described above. Due to fermentation by the strains, in addition to succinic acid, other dissociated organic acids (e.g., matric acid, citric acid, oxal acetic acid, formic acid, acetic acid, fumaric acid, etc.), cations (K ⁺ , NH4 ⁺ , Mg ^{2 +} , Na ⁺ ) , anions _{^{(CO 3 2 -, PO 4}} -, SO 4 2 -), ( hereinafter referred to as "succinic acid fermentation broth expression ') unreacted carbohydrate fermentation broth comprises an amino acid with an excess of water is formed.

In this way, the base 40 is added to the fermentation broth in which the succinic acid is expressed while the fermentation is in progress. At this time, the base 40 to be added is not particularly limited as long as it can selectively precipitate succinic acid in the fermentation broth, similarly to the first method of forming the precipitate containing the succinate described above. Examples of such materials include, but are not limited to, calcium hydroxide (Ca (OH) ₂ ), magnesium hydroxide (Mg (OH) ₂ ), calcium carbonate (CaCO ₃ ), magnesium carbonate (MgCO ₃ ), and mixtures thereof. .

Here, among the succinates formed by reacting the base with dissociated succinic acid (succinic anion), calcium succinate (solubility: about 15 g / l) is more precipitated than magnesium succinate (solubility: about 45 g / l). In this case, it is preferable to use calcium hydroxide or calcium carbonate. Meanwhile, in order to prepare succinic acid using a strain, carbonate ions must be sufficiently supplied as a carbon source to be dissolved in the fermentation broth. Dissolution is advantageous for fermentation due to high feeding efficiency. As such, it may be appropriate to use calcium carbonate or magnesium carbonate as a source of carbon ions and capable of forming precipitate salts. More preferably, it is appropriate to use calcium carbonate which can be fed at a temperature of carbonate and which can selectively precipitate succinic acid.

When such a base 40 is added to the fermentation broth in which succinic acid is expressed, succinic acid in the fermentation broth causes an acid-base reaction with the base to form succinate. For example, when the added base is calcium hydroxide, calcium succinate may be formed, and when the added base is magnesium hydroxide, magnesium succinate may be formed.

At this time, the succinate formed is precipitated because the solubility at room temperature is lower than that of succinic acid in the fermentation broth (solubility: about 78 g / l). For example, when the succinate formed is calcium succinate, it is precipitated because the solubility of the calcium succinate at room temperature is lower than that of succinic acid in the fermentation broth at about 15 g / l. When the succinate formed is magnesium succinate, it is precipitated because its solubility is about 45 g / l at room temperature, which is lower than that of succinic acid. Since the precipitate produced during the fermentation does not affect the growth of the strain, succinic acid can be continuously produced without product inhibition, thereby producing a high concentration of succinic acid.

On the other hand, the fermentation by the strain may be different from the pH conditions for optimal fermentation depending on the type of strain, in general, the optimum fermentation may be made at about pH 5 to 7. However, due to the production of succinic acid during fermentation, the pH of the fermentation broth gradually decreases to about 3 to 4, and thus, fermentation may not be performed properly. By appropriately adding a base in powder form or slurry form to such fermentation broth, the pH of the fermentation broth in which the succinic acid is expressed can be maintained at about 5-7.

In addition, when the acid when expressed and adding a base to the fermentation broth with, the salt, i.e., succinate derived from succinic acid in the fermentation broth is selectively precipitated, the fermentation broth in the anion _{^{(CO 3 2-, PO 4 -}} , SO 4 2 ^- ), Like succinic acid, can be precipitated by reaction with added base. In particular, in the case of carbonate ion used as a carbon source during fermentation by the strain, it is reacted with the base added to the fermentation broth because it is continuously supplied in the form of carbon dioxide gas or carbonate during fermentation. Due to the reaction of the carbonate with the base, a considerable amount of precipitated carbonates such as calcium carbonate salts and magnesium carbonate salts are formed and precipitated together with succinate salts, which in turn leads to a decrease in purity of the precipitate containing succinate salts. In addition, although phosphate or sulfate ions, which are essential components for the growth of the strain, are contained in trace amounts in the fermentation broth, if these ions are precipitated by reacting with the added base, the fermentation of the strain may be inhibited.

Therefore, in the present invention, the pH of the fermentation broth expressing succinic acid was maintained at about 4.5 to 6.5 or less upon addition of the base so that more precipitates of succinate were formed and less precipitates of salts induced by anions were formed. This is because the reaction of the succinic acid anion-base has a higher selectivity than the reaction of the other anion-base at a low pH. Thereby, the purity of the precipitate containing succinate produced by minimizing the formation of salts induced by anions can be improved.

As described above, the formation of a precipitate containing succinate by adding a base during fermentation may be performed during the fermentation process in the form of batch, fed-batch, or continuous form.

For example, when the step of forming a precipitate containing succinate by adding a base during fermentation by the strain is carried out in the form of Batch, Fed-batch, the concentration of the precipitate containing succinate is about 10 to After stopping the fermentation at 30% by weight, the precipitate containing the succinate produced using a decanter, filter, or centrifuge is separated from the first supernatant, recovered and later transferred to the succinic acid crystal forming region. do. On the other hand, when the step of forming a precipitate containing succinate by adding a base during the fermentation by the strain is carried out in a continuous fermentation form, the succinate is produced in the rear end of the fermenter using a decanter, a filter, or a centrifuge The precipitate is separated from the first supernatant, recovered and then transferred to the succinic acid crystal region.

In this case, the used strain 20 may be easily separated from the precipitate 60 containing the succinate because the strain is suspended in the first supernatant 50. As such, the first supernatant 50 containing the strain may be recovered and reused when the fermentation broth is formed.

However, when the separated first supernatant 50 is recovered and reused, when the concentration of impurities in the first supernatant reaches a level that inhibits the strain fermentation, the impurities in the first supernatant are purged through a purge process. It is appropriate to reuse after reducing the concentration to below a certain level.

Specifically, when the concentration of impurities in the separated first supernatant is at a level inhibited by strain fermentation in the range of about 1 wt% or more, preferably 1 wt% to 2 wt%, the first supernatant 50 Is added directly to the purge zone (IV) rather than directly used to form the fermentation broth, and the amount of water is discarded and the amount of impurities in the first supernatant is reduced to 1 wt% or less. After making it suitable for reuse in the formation of fermentation broth.

II. Formation of succinic acid in crystalline state by addition of acidic aqueous solution

The precipitate 60 containing the succinate obtained by the above-mentioned methods is transferred to the succinic acid crystal forming region (II), as shown in Figs. 1 to 3, into which an aqueous hydrochloric acid solution and / or an aqueous nitric acid solution ( 70) is fed (added) so that the succinate in the precipitate 60 containing the succinate reacts with aqueous hydrochloric acid solution and / or aqueous nitric acid solution to produce succinic acid as a reaction product, and calcium chloride and / or calcium nitrate as by-products. Create In addition, salts induced by anions contained in the precipitate together with succinate salts and salts derived from other organic acids include, for example, calcium sulfate, calcium phosphate, calcium carbonate, calcium organic acid salts, and / or aqueous solution of nitric acid. The reaction produces impurities such as sulfuric acid, phosphoric acid, carbonic acid, and other organic acids.

However, the succinic acid 90 produced is precipitated because of its relatively low solubility as compared to the by-products (calcium chloride and / or calcium nitrate) or the impurities. On the other hand, by-products or impurities having relatively high solubility are dissolved in the supernatant (hereinafter referred to as 'second supernatant') 80 and contained in the second supernatant. For example, when an aqueous hydrochloric acid solution 70 is added to the precipitate 60 containing calcium succinate, the calcium succinate in the precipitate reacts with the aqueous hydrochloric acid solution to produce a byproduct calcium chloride along with the reaction product succinic acid, and also in the precipitate. Salts derived from anions and salts derived from other organic acids react with aqueous hydrochloric acid to produce impurities such as sulfuric acid, phosphoric acid, carbonic acid and other organic acids. At this time, the resulting succinic acid (solubility: about 78 g / l) is relatively low solubility compared to by-product calcium chloride (solubility: about 745 g / l) or impurities, succinic acid is precipitated without dissolving in the second supernatant, Calcium chloride or impurities are dissolved in the second supernatant and present in the second supernatant.

As such, when the aqueous hydrochloric acid solution and / or the aqueous nitric acid solution 70 is added to the precipitate 60 containing the succinic acid salt, calcium chloride and / or have a higher solubility than succinic acid together with succinic acid. Or since calcium nitrate is formed, only succinic acid can be easily obtained in the crystalline state. Therefore, unlike the case of using a conventional aqueous sulfuric acid solution, in order to separate the succinic acid, there is no need for additionally obtaining a succinic acid in crystalline state through water evaporation, and only using a low-cost filtration device such as a filter to obtain succinic acid in the crystalline state. You can get it.

In the present invention, the succinate-containing precipitate 60 and the aqueous hydrochloric acid solution (and / or nitric acid solution) 70 are suitably mixed in such a ratio that the succinate precipitate can be replaced with succinic acid crystals. If the use ratio of aqueous hydrochloric acid solution (and / or nitric acid solution) is too small, the substitution with succinic acid may not occur completely, and if the use ratio of aqueous hydrochloric acid solution (and / or nitric acid solution) is too high, the second supernatant Hydrochloric acid (and / or nitric acid) remains in the process, resulting in additional costs for recycling the second supernatant. Therefore, at least one acidic solution in the precipitate containing succinate, aqueous solution of hydrochloric acid and aqueous solution of nitric acid is succinate in the precipitate containing succinate (a): acid (b) in acidic aqueous solution = 1: 1-1: 5 Preference is given to using in molar equivalents.

Further, in the present invention, it is important to use an aqueous solution of hydrochloric acid and / or an aqueous solution of nitric acid in order to minimize the amount of succinic acid dissolved and lost in the second supernatant later and to minimize impurities in the succinic acid crystals. For example, when an excessively diluted aqueous hydrochloric acid solution and / or an aqueous nitric acid solution is used, the amount of succinic acid dissolved and lost in the supernatant is increased to lower the separation yield. On the other hand, when using excessively concentrated aqueous hydrochloric acid and / or aqueous nitric acid solution, impurities (particularly calcium chloride and / or calcium nitrate) may be included in the succinic acid crystals to reduce the quality of the final succinic acid as well as the reaction product. It may also impair fluidity. Thus, in the present invention, an aqueous hydrochloric acid solution having a concentration of about 10 to 40% by weight, preferably about 20 to 35% by weight is used, and / or the concentration is about 20 to 70% by weight, preferably about 30 to 60% by weight. A nitric acid aqueous solution of% was used.

The acidic aqueous solution 70 of at least one of the aqueous hydrochloric acid solution and nitric acid solution is suitably added to the precipitate 60 containing the succinate salt at room temperature or at a high temperature of about 50 to 100 ° C.

On the other hand, as a result of the reaction of the precipitate 60 containing the succinate and the aqueous hydrochloric acid solution and / or the aqueous nitric acid solution 70, the second supernatant 80 formed together with the succinic acid 90 in the crystalline state is separated from the succinic acid in the crystalline state. Afterwards, the acid 70 and base 40 may be separated, recovered and reused, optionally via a hydration reaction at high temperature. Specifically, the second supernatant 80 separated from the succinic acid in the crystalline state in the hydration reaction zone (V) is contained in the second supernatant at a temperature in the range of about 400 to 800 ° C., preferably about 400 to 600 ° C. Salts, such as calcium chloride or calcium nitrate, may be converted to hydrochloric acid or base 40 such as nitric acid 70 and calcium hydroxide. At this time, the generated hydrochloric acid or nitric acid 70 can be recovered and reused in the step (II) of producing succinic acid in a crystalline state, while the base 40 such as calcium hydroxide produced is recovered to contain the succinate-containing precipitate. It can be reused in the process (I) to produce a. However, calcium chloride or calcium nitrate contained in the separated second supernatant may be recovered and used by itself.

III. High purity succinic acid formation

Meanwhile, in the present invention, the succinic acid 90 in the crystalline state may be selectively washed in the washing region (III) using a saturated aqueous succinic acid solution 100 (pH about 2.5, 7% succinic acid solution). As a result, a small amount of impurities (CaCl ₂ , CaCO ₃ , CaSO _4, etc.) buried in the succinic acid 90 in the crystalline state may be removed, thereby obtaining succinic acid 120 having a more improved quality. Thereafter, the saturated succinic acid solution 110 containing impurities contained in the succinic acid in the crystal state may be selectively transferred to the ion exchange resin region (VI), and then the saturated aqueous succinic acid solution may be purified using an ion exchange resin. In addition, the purified saturated succinic acid solution 100 may be recovered and used again in the washing region (III).

Succinic acid isolated and purified at high efficiency and low cost by the method described above is not only used in the food, pharmaceutical and cosmetic industries currently used, but also BDO (1,4-butanediol), THF (tetrahydrofuran) and GBL (gamma-butyrolactone) , NMP (n-methylpyrrolidone) and the like can be used as a raw material for the production of C4 general-purpose products, it can be used in various other fields.

Hereinafter, the present invention will be described in more detail by way of examples. However, an Example is for illustrating this invention and is not limited only to these.

[ Example  One]

Example 1-1 Formation of Precipitate Containing Succinate

The succinic acid-expressing fermentation broth (concentration of succinic acid in the fermentation broth: 79.2 g / l) was removed using a centrifuge (12,000 rpm, 10 minutes). Thereafter, 300 ml of the fermentation broth from which the strain was removed was heated to a temperature of 80 ° C. while stirring to remove carbonate ions from the fermentation broth.

To 300 ml of the fermentation broth from which the carbonate was removed, 16.24 g of calcium hydroxide (Ca (OH) ₂ ) was added under stirring to bring the pH of the fermentation broth to about 7.0, which was then cooled to room temperature, using a filter (0.6 μm). Filtration resulted in a precipitate and a supernatant.

Here, the concentration of calcium succinate in the supernatant was analyzed by HPLC (LC-20AD Prominence Series With Standard Accessories, SHIMADZU), and the concentration of calcium succinate in the supernatant was about 11 g / l. This means that about 88% by weight of succinic acid was precipitated in the form of calcium succinate.

Experimental Example 1

In order to confirm the purity of the precipitate containing succinate obtained according to the present invention, the following analysis was performed using the precipitate obtained in Example 1-1 and the supernatant.

1) Organic Acid Analysis

HPLC (LC-20AD Prominence Series With Standard Accessories, SHIMADZU) was used to analyze the fermentation broth before calcium hydroxide addition and the precipitates obtained after calcium hydroxide addition and the organic acids contained in the supernatant, respectively. At this time, 1 ml of the supernatant and 1 g of the precipitate were diluted 100-fold and analyzed, respectively, and the analysis results are shown in Table 1 below.

ingredient Fermentation broth before adding calcium hydroxide (% by weight) ^a After adding calcium hydroxide Supernatant (% by weight) ^a Sediment (wt%) ^a Succinic acid (salt) 99.18 96.00 99.86 Pyruvic acid (salt) 0.44 3.25 0.03 Citric Acid (salt) 0.35 0.57 0.10 Oxalic acid (salt) 0.03 0.18 0.02 ( ^a : weight% of each component with respect to the total dry weight)

As can be seen in Table 1, the ratio of calcium succinate in the precipitate formed after adding calcium hydroxide was higher than the ratio of calcium succinate in the supernatant. Therefore, it was found that the addition of a base such as calcium hydroxide to the fermentation broth expressing succinic acid can selectively precipitate the succinic acid salt as compared to other organic acid induced precipitation salts.

2) ion analysis

ICP (720-ES. Varian), and AA (AAnalyst 300, PERKIN ELMER) was used to analyze the ions contained in the precipitate and the supernatant obtained in Example 1-1, respectively, and the results are shown in Table 2 below. . At this time, 1 ml of the supernatant was diluted 1,000-fold, and 1 g of the precipitate was diluted 10,000-fold and analyzed.

Ionic component Content in Supernatant (%) Content in Sediment (%) P 7.8 92.2 S 35.2 64.8 Na 86.1 13.9 K 89.4 10.6 Mg 83.5 16.5 Ca 9.5 90.5

As can be seen in Table 2, when the pH is to adjust the salt amount to be 7, the cation (K ^+, Mg ^+, Na ^+), while mainly present in the supernatant, anions (CO ₃ ^{2 -,} PO ₄ ^-, SO ₄ ^2-) was confirmed that it is included in the colored precipitate in the form of calcium sulfate, calcium phosphate, calcium carbonate reacts with calcium hydroxide.

Experimental Example 2

According to the present invention, when a base was added to a fermentation broth expressing succinic acid to form a precipitate containing succinate, it was observed whether or not a precipitate containing succinate was formed in order to confirm the effect of the addition amount of the base. In addition, the concentration of succinate in the resulting supernatant was measured using HPLC, and the ratio of each ion contained in the precipitate and the supernatant was analyzed using IPC and AA.

At this time, the precipitate and the supernatant were obtained in the same manner as in Example 1-1 except that calcium hydroxide was added to 300 ml of the fermentation broth from which the carbonate was removed so that the pH of the fermentation broth was 4.1, 5.0, 6.3, and 7.0.

1) As a result of the observation, when calcium hydroxide was added so that the pH of the fermentation broth was 4.1, no precipitate could be observed. On the other hand, when calcium hydroxide was added so that the pH of the fermentation broth was 5.0 or more, a precipitate could be observed. Thus, in the present invention, it was found that it is appropriate to add the base to the fermentation broth expressed succinic acid so that the pH of the fermentation broth is in the range of more than 4.1 to 7.0 or less.

2) As a result of analyzing the concentration of succinate in each supernatant obtained by HPLC, the supernatant obtained by adding a base such that the pH of the fermentation broth was 5.0 contained succinate at a concentration of about 30 g / l. The supernatant obtained by adding a base to the pH of 6.3 contained succinate at a concentration of about 17 g / l, and the supernatant obtained by adding a base such that the pH of the fermentation broth was 7.0 was about 11 g / l. It was contained at the concentration of.

3) In addition, the results of analyzing the ratio of ions contained in each precipitate and the supernatant using ICP and AA are shown in Table 3 below.

% Content of ions in the supernatant pH 5.0 pH 6.3 pH 7.0 P 84.6 20.7 7.8 S 58.2 44.1 35.2 Na 84.0 82.8 86.1 K 84.2 83.4 89.4 Mg 83.8 80.9 83.5 Ca 43.9 25.1 9.5

As can be seen in Table 3, when the pH of the fermentation broth is adjusted by the amount of base added, the cation (K ⁺ , Mg ⁺ , Na ⁺ ) is mainly present in the supernatant regardless of the pH of the fermentation broth, while _{^{(CO 3 2 -, PO 4}} -, SO 4 2-) increased the rate at which increasing pH contained in the precipitate in the form of calcium sulfate, calcium phosphate, calcium carbonate. From these results, when a base such as calcium hydroxide is added to the fermentation broth to which succinic acid is expressed to form a succinate precipitate, the lower the pH of the fermentation broth, the more succinate precipitate can be precipitated more selectively than the anion-induced salt precipitate. I knew it was.

[Example 1-2]

To 12 g of the precipitate obtained in Example 1-1, 1 equivalent (16.0 g) of 35% aqueous hydrochloric acid solution and 10.4 ml of distilled water were added, followed by stirring to react the precipitate with aqueous hydrochloric acid solution. The precipitate and the supernatant obtained after the reaction were obtained by separating the precipitate and the supernatant using a filter (0.6 μm).

At this time, the concentration of succinic acid in the supernatant obtained above was analyzed by HPLC, and the concentration of succinic acid was about 26 g / l. This means that 96% of succinic acid was obtained from the precipitate (precipitate succinate).

[Example 2]

Example 1-1, except that 1 equivalent (16.0 g) of 35% aqueous hydrochloric acid solution and 2 equivalents (32.0 g) of aqueous 35% hydrochloric acid solution were added instead of 10.4 mL of distilled water to 12 g of the precipitate obtained in Example 1-1. The precipitate and the supernatant were obtained in the same manner as in -2.

At this time, the concentration of succinic acid in the supernatant obtained above was analyzed by HPLC, and the concentration of succinic acid was about 26 g / l. This means that 96% of succinic acid was obtained from the precipitate (precipitate succinate).

Example 3

To 12 g of the precipitate obtained in Example 1-1, except that 1 equivalent (16.0 g) of 35% aqueous hydrochloric acid solution and 10.4 ml of distilled water were added 1 equivalent (16.0 g) of 60% nitric acid solution and 14.3 ml of distilled water. In the same manner as in Example 1-2, a precipitate and a supernatant were obtained.

At this time, the concentration of succinic acid in the supernatant obtained above was analyzed by HPLC, and the concentration of succinic acid was about 31 g / l. This means that 95% of succinic acid was obtained from the precipitate (precipitate succinate).

[Experimental Example 3]

In order to determine the purity of the succinic acid of the crystal state obtained according to the present invention, the following analysis was carried out using the precipitates prepared in Examples 1-2, 2 and 3 and the supernatant.

1) Organic Acid Analysis

HPLC (LC-20AD Prominence Series With Standard Accessories, SHIMADZU) was used to analyze precipitates before addition of aqueous hydrochloric acid solution or nitric acid solution, and precipitates obtained after addition of aqueous hydrochloric acid solution or nitric acid solution, and organic acids contained in the supernatant, respectively. At this time, 1 ml of each supernatant obtained by Examples 1-2, 2 and 3 and 1 g of precipitate were diluted 100-fold and analyzed, respectively, and the analysis results are shown in Table 1 below.

Sediment before adding hydrochloric acid
(Wt%) ^a Example 1-2 Example 2 Example 3 Supernatant precipitate Supernatant precipitate Supernatant precipitate Suche mountain 99.86 98.63 99.98 99.17 99.98 99.02 99.97 Pyruvic acid 0.03 0.22 0.002 0.16 0.002 0.15 0.005 Citric acid 0.10 1.13 0.010 0.65 0.008 0.79 0.011 Oxal acetic acid 0.02 0.02 0.007 0.02 0.008 0.03 0.009 ( ^a : weight% of each component with respect to the total dry weight)

As can be seen in Table 4, the ratio of succinic acid contained in the precipitate obtained by the addition of aqueous hydrochloric acid solution or aqueous nitric acid solution was higher than the ratio of succinic acid contained in the supernatant. It is derived from calcium succinate, other organic acid derivatives and anion derivatives (calcium sulphate, calcium phosphate, calcium carbonate) in the precipitate after the reaction of the precipitate with aqueous hydrochloric acid or aqueous nitric acid solution. ), Which is considered to be because the products other than succinic acid have solubility in comparison with succinic acid and mainly exist in aqueous solution. In addition, the solubility of calcium chloride and calcium nitrate was 745 g / l and 1212 g / l, respectively, which is very high compared to the succinic acid solubility. Furthermore, the theoretical solubility of succinic acid is about 78 g / l, Solubility was as low as 26 g / l and 31 g / l, respectively. This is because organic acids tend to exist in the form of non-lactic acid when pH is lowered in general, and it is thought that the organic acid monomers are bonded to each other due to hydrogen bonding of monomer organic acids, and thus the crystal size increases.

From this, it was found that when succinic acid was formed by adding an aqueous hydrochloric acid solution or an aqueous nitric acid solution to the precipitated succinate salt, succinic acid could be selectively precipitated compared to other organic acids.

2) ion analysis

ICP (720-ES. Varian) and AA (AAnalyst 300, PERKIN ELMER) were used to analyze the ions contained in the precipitates and supernatants obtained in Examples 1-2, 2 and 3, and the results are shown in the following table. 5 is shown. At this time, 1 ml of each supernatant was diluted 1,000-fold, and 1 g of each precipitate was diluted 10,000-fold and analyzed.

% Content of ions in the supernatant Example 1-2 (% by weight) ^a Example 2 (% by weight) ^a Example 3 (% by weight) ^a P 82.19 89.03 87.45 S 82.31 84.79 82.05 Na 73.75 78.90 77.92 K 70.79 87.81 80.13 Mg 84.38 97.27 97.20 Ca 87.72 90.11 87.92 ( ^a : weight% of each component with respect to the total dry weight)

As a result of the analysis, in each of the supernatants obtained in Examples 1-2, 2 and 3, salts (eg calcium sulfate, calcium phosphate, calcium carbonate) induced by anions contained in the precipitate were converted into sulfuric acid, phosphoric acid and carbonic acid. It was found that it contained a large amount.

From these results, it can be seen that when succinic acid containing an aqueous solution of hydrochloric acid or nitric acid is added to the succinate-containing precipitate to form crystalline succinic acid, succinic acid may be selectively precipitated compared to acids to which salts induced by anions are converted. Could.

Thus, the succinic acid separation and purification method of the present invention, unlike the conventional succinic acid separation and purification method, it was confirmed that the succinic acid crystals can be easily obtained by using a filter such as a filter.

Comparative Example 1

A precipitate and a supernatant were obtained in the same manner as in Example 1-2, except that 1 equivalent (7.5 g) of 98% sulfuric acid aqueous solution and 20.8 ml of distilled water were added to 12 g of the precipitate obtained in Example 1-1.

In the case of the precipitates and supernatants obtained in Examples 1-2, 2 and 3, in contrast to the precipitates and supernatants obtained in Comparative Example 1, the succinic acid was present in the precipitate and calcium chloride was present in the supernatant to separate the succinic acid. Calcium sulfate (solubility: 2.4 g / l) was precipitated together and it was difficult to separate only succinic acid. From this, it was found that when a high concentration of sulfuric acid solution is used, calcium sulfate precipitates together with succinic acid, making it difficult to selectively separate succinic acid.

Example 4

6 g of the precipitate obtained in Example 2 was washed with 20 ml of saturated succinic acid solution (concentration: 7% by weight).

[Experimental Example 4]

In order to know the degree of succinic acid contained in the precipitate obtained in Example 4 and the supernatant, HPLC (LC-20AD Prominence Series With Standard Accessories, SHIMADZU) was used to wash the precipitate before the aqueous saturated succinic acid solution and the saturated aqueous succinic acid solution. The organic acid contained in each of the obtained precipitate and the supernatant was analyzed. The results are shown in Table 6 below. At this time, 1 ml of the supernatant and 1 g of the precipitate were diluted 100-fold and analyzed.

ingredient Precipitation before washing (% by weight) ^a Sediment after washing Supernatant (% by weight) ^a Sediment (wt%) ^a Suche mountain 99.98 99.966 99.9977 Pyruvic acid 0.002 0.003 0.0004 Citric acid 0.008 0.001 0.0010 Oxal acetic acid 0.008 0.029 0.0007 ( ^a : weight% of each component with respect to the total dry weight)

As can be seen in Table 6, by washing the precipitate using a saturated aqueous succinic acid solution, it was found that traces of organic acid impurities and salts present in the precipitate were removed, and as a result, high purity succinic acid of 99.998% was obtained. there was.

1 is a process chart for separating and purifying succinic acid from a fermentation broth according to one embodiment of the present invention.

2 is a process chart for separating and purifying succinic acid from a fermentation broth according to another embodiment of the present invention.

3 is a process chart for separating and purifying succinic acid from a fermentation broth according to another embodiment of the present invention.

<Explanation of symbols for the main parts of the drawings>

I-1 (a): fermentation broth production zone, 1-1 (b): succinate formation zone,

I-2: fermentation broth and succinate forming region,

II: succinic acid crystal forming region, III: washing region,

IV: purge zone, V: hydration zone,

Ⅵ: Ion exchange resin region 10: Carbohydrate, CO ₂ , nutrients, etc.

20: strain, 30: fermentation broth expressing succinic acid,

40: base, 50: first supernatant,

60: precipitate containing succinate, 70: aqueous hydrochloric acid (nitric acid) solution,

80: second supernatant, 90: succinic acid in crystalline state,

100: saturated succinic acid solution, 110: impurity-containing saturated succinic acid solution,

120: high purity succinic acid

Claims (16)

As a method for separating and purifying succinic acid from a fermentation broth prepared with succinic acid using a strain, Generating a precipitate containing succinate by adding a base to a fermentation broth in which succinic acid is expressed or expressed using a strain; And Generating an crystalline succinic acid by adding an acidic aqueous solution selected from aqueous hydrochloric acid, aqueous nitric acid and a mixture thereof to the precipitate containing succinate. Succinic acid separation and purification method characterized in that it comprises a. The method of claim 1, wherein the pH of the fermentation broth expressed or expressed succinic acid is maintained in the range of 4.5 to 6.5 when the base is added. The method of claim 1, wherein the succinic acid is expressed or expressed using the strain is characterized in that the succinic acid isolation and purification method characterized in that the batch form, fed-batch form, or in a continuous form. The succinic acid of claim 1, wherein the base is selected from the group consisting of calcium hydroxide (Ca (OH) ₂ ), magnesium hydroxide (Mg (OH) 2), calcium carbonate (CaCO ₃ ) and magnesium carbonate (MgCO ₃ ). Separation and Purification Methods. The method of claim 1, wherein the addition of the base to the succinic acid-expressed fermentation broth is characterized in that carried out at room temperature or a temperature of 50 to 100 ℃. According to claim 1, Before adding the base to the fermentation broth expressed succinic acid, Succinic acid separation and purification method characterized in that it further comprises the step of removing the carbonate (CO ₃ ^2- ) contained in the fermentation broth by heating the fermentation broth expressing the succinic acid or by supplying air or nitrogen. The method of claim 6, wherein the heating temperature of the fermentation broth is characterized in that 50 to 100 ℃ succinic acid separation and purification method. The method of claim 1, wherein in the step of producing a succinate-containing precipitate, the first supernatant obtained in addition to the succinate-containing precipitate is recovered and reused in the formation of the fermentation broth expressing the succinic acid. Purification method. The succinic acid separation of claim 1, further comprising maintaining the concentration of impurities in the first supernatant in a range of 1 wt% or less when the concentration of the impurities in the first supernatant is greater than 1 wt%. And purification methods. According to claim 1, wherein the concentration of the hydrochloric acid aqueous solution is in the range of 10 to 40% by weight, Succinic acid separation and purification method characterized in that the concentration of the aqueous nitric acid ranges from 20 to 70% by weight. The method according to claim 1, wherein the succinate (a) contained in the succinate-containing succinate and the acid (b) in the acidic aqueous solution are used in a ratio of a: b = 1: 1 to 1: 5 molar equivalents. Characterized by succinic acid separation and purification method. The method of claim 1, wherein the addition of the acidic aqueous solution is carried out at room temperature or at a temperature of 50 to 100 ℃ succinic acid separation and purification method. According to claim 1, Succinic acid characterized in that it further comprises the step of producing an acid and a base at a temperature in the range of 400 to 800 ℃ the second supernatant produced in the crystalline succinic acid production step Separation and Purification Methods. The method of claim 13, wherein the generated base is recovered and reused in the production of a precipitate containing succinate, Succinic acid separation and purification method, characterized in that for recovering the generated acid and reuse when generating the succinic acid in the crystal state. According to claim 1, After the succinic acid production step of the crystalline state, Succinic acid separation and purification method characterized in that it further comprises the step of washing the succinic acid of the crystalline state produced in the succinic acid production step using a saturated aqueous succinic acid solution. The succinic acid separation and purification method according to claim 15, wherein the saturated succinic acid solution used in the washing step is recovered, purified by ion exchange resin, and the purified saturated succinic acid solution is reused in the washing step.

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